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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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D) Molecular Cartography localizing distinct CD45 cells. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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E) Circos plot showing NicheNet predicted ligand-receptor pairs between KCs and LSECs, HSCs and hepatocytes uniquely expressed in the human liver (left) and UMAPs showing normalized expression of indicated chemokines in the human liver (right) and CCR1/Ccr1 in the human and murine NAFLD liver (bottom). (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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F) Zonation of indicated genes in the healthy and steatotic human liver. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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G) UMAP of murine NAFLD (SD and WD) stromal cells (4,025 cells/nuclei) isolated from Figure S5J and re-clustered with scVI (left) and proportions of each cell type in SD- and WD-fed mice (right). (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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H) Top DEGs between cell types. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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I) Mice were fed a western diet (WD) or standard diet (SD) for 36 weeks to induce NAFLD, and Visium analysis was performed.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Analysis is pooled from 1 liver slice from the SD condition and 3 liver slices from the WD condition.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Zonation of Ccl2 fibroblasts and fibroblasts in SD- and WD-fed mice.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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See also Tables S3 and S10.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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We next aimed to investigate the role of CD45 cells in the regulation of LAM location in the healthy versus steatotic liver.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Given the low number of steatotic human samples in our sc/snRNA-seq analysis and the variation between patients, we turned to the murine NASH model to investigate this.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Zooming in on SCs (Figure 6G; Table S10), we identified that there was an increase in fibroblasts in the WD-fed mice.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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There was also a considerable overlap between fibroblasts and HSCs in terms of their gene expression profiles suggesting there may be a differentiation trajectory between the HSCs and the fibroblasts, although this remains to be validated in vivo.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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We also noted a sub-population of fibroblasts expressing Ccl2, a known ligand for CCR2 which is expressed on monocytes recruited to the liver during NAFLD (Remmerie et al., 2020), as well as Cd44 and Vcam1, two genes involved in monocyte recruitment and adhesion (Johnson and Ruffell, 2009; Meerschaert and Furie, 1995).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Both fibroblast populations were enriched in the periportal steatotic regions (Figure 6I), in which we also observe enrichment of the LAMs (Figure 5I).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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We also found high CCL2 and Cd44 expression in human fibroblasts (Table S3), potentially recruiting bile-duct LAMs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Together, this suggests that fibroblasts may play an important role in recruiting LAMs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Having identified a potential role for the mac niche cells in regulating mac subset location, we next sought to determine the involvement of these cells in regulating mac phenotypes.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To assess the roles of conserved cell-cell interactions in driving mac heterogeneity across species, we performed a differential NicheNet (Browaeys et al., 2019) analysis between the distinct hepatic macs and the CD45 cells present in their respective niches focusing on ligands and receptors conserved in both human and mouse.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This revealed very few specific ligand-receptor pairs for LAMs (Figure S8G; data not shown), hinting that local factors such as metabolites rather than unique cell-cell interactions may drive the LAM phenotype.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Indeed, this would be consistent with the presence of LAM-like cells in multiple tissues including the obese adipose tissue, the brain, the lung, and the heart (Jaitin et al., 2019; Keren-Shaul et al., 2017; Liao et al., 2020; Rizzo et al., 2020).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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In line with this, BM monocytes cultured with acetylated low-density lipoprotein expressed LAM-associated genes (Figure 7A), demonstrating a dominant role for lipids in inducing the LAM phenotype.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Conversely, for KCs, we found multiple ligand-receptor pairs to be conserved between human and mouse (Figures 7B and S8G).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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One of these, an activin receptor-like kinase (ALK1)-bone morphogenic protein (BMP)9/10 circuit between KCs (ALK1; encoded by Acvrl1) and stellate cells (BMP)9/10 encoded by Gdf2/Bmp10 respectively) was found to be conserved in all 7 species and was predicted to control the expression of a number of the conserved KC genes (Figures 7B, 7C, and S8H).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To validate a role for this axis in KCs, we generated Fcgr1-Cre × Acvrl1 mice, eliminating ALK1 specifically from CD64-expressing macs (Scott et al., 2018).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This led to an almost complete loss of VSIG4 KCs (Figures 8D–8F), demonstrating that evolutionarily conserved ALK1 signaling is crucial for KCs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To determine if ALK1 is required for KC maintenance we generated Clec4f-Cre × Acvrl1 mice, eliminating ALK1 only from differentiated KCs (Scott et al., 2018).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This revealed a relatively similar phenotype than observed in the Fcgr1-Cre mice, suggesting that ALK1 is also required for KC maintenance (Figure S8I).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To directly test the need for ALK1 in KC development, we generated BM chimeras whereby CD45.1 Clec4f-Dtr mice were irradiated with their livers shielded to avoid any radio damage.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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These mice were then reconstituted with either CD45.2 Acvrl1 or Fcgr1-CrexAcvrl1 BM.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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4 weeks later, mice were given a single i.p.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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injection of DT to deplete the KCs and 7 or 13 days thereafter chimerism within the KC population was determined (Figure 7G).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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We observed that, already by day 7, KO BM-derived cells were almost completely outcompeted by WT BM-derived cells within the KC pool.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This was not observed in monocytes but a similar pattern was observed in VSIG4 macs demonstrating that ALK1 is critically required for early KC development (Figure 7H).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Finally, while NicheNet predicts that BMP9/10 from stellate cells would signal through ALK1 to induce KC development and maintenance, a prediction in line with our previous NicheNet analysis (Bonnardel et al., 2019), another recent study has suggested that transforming growth factor (TGF)β signaling would be important for KCs (Sakai et al., 2019).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This prediction was made on the basis of SMAD4 signaling, which is a common downstream effector of both TGF-β Receptor and ALK1-induced signaling.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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As TGF-β was also found to participate in an ALK1-TGF-β receptor containing signaling complex (Goumans et al., 2003), we thus examined whether TGF-β signaling is important for KCs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To this end we utilized an ALK1-Fc trap or TGF-β type II receptor (TGFβRII)-Fc trap alongside appropriate isotype controls.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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ALK1-Fc selectively sequesters BMP9/10 (Desroches-Castan et al., 2021), while TGFβRII-Fc selectively interferes with TGF-β1/TGF-β3 receptor binding (Komesli et al., 2017).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Clec4f-Dtr mice were depleted of KCs and simultaneously treated with either receptor-Fc traps or isotype controls and KC development was examined 7 days later (Figure 7I).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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In line with the chimera study, treatment with ALK1-Fc significantly abrogated KC development; however, blocking TGFβ signaling had only a minor effect on the proportion of VSIG4 macs (Figure 7J).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Altogether this validates the NicheNet prediction and demonstrates that the evolutionarily conserved ALK1-BMP9/10 axis is crucial for the development and maintenance of KCs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Figure 7ALK1-BMP9/10 axis regulates KC development(A) Mouse BM monocytes were cultured in the presence of CSF1 and indicated concentrations of human ac-LDL, prior to analyzed for expression of indicated genes by qPCR.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 2 experiments.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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One-way ANOVA with Bonferroni post-test compared with 0 ng/mL.(B) NicheNet circos plot highlighting conserved ligand-receptor pairs and induced target genes between KCs and indicated niche cells in human and mouse.(C) Feature plots showing expression of ALK1 (Acvrl1) in human myeloid cells (left) and GDF2/BMP10 in CD45 cells (right).(D) Livers were harvested from Fcgr1-CrexAcvrl1 mice or Acvrl1 or Acvrl1 controls and KCs examined (left) and quantified (right) using VSIG4 expression.(E) Expression of indicated KC markers by mac populations in Fcgr1-CrexAcvrl1 or Acvrl control mice.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 3 independent experiments with n = 9 per group.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Student’s t test.(F) Expression of indicated markers in livers of Fcgr1-CrexAcvrl1 or Acvrl1 or Acvrl1 control mice by confocal microscopy.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Scale bars, 50 μm.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Images are representative of 2 mice per group.(G) Schematic of chimera experiment setup.(H) % chimerism normalized to levels in blood Ly6C monocytes in Clec4f-Dtr mice 7 or 13 days after DT administration following partial irradiation and receiving either Acvrl or Fcgr1-CrexAcvrl BM 4 weeks earlier.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 2 independent experiments with n = 10–12 mice per group.(I) Schematic of Fc trap experiment setup.(J) Representative FACS plots showing VSIG4 and CLEC2 expression by total macs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Numbers represent % of total mac population in the indicated gate (left) and % of VSIG4 and VSIG4 macs among total CD45 cells in the different treatment conditions.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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One-way ANOVA with Bonferroni post-test.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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p < 0.05, p < 0.01, p < 0.001, p < 0.0001.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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See also Figure S8.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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ALK1-BMP9/10 axis regulates KC development (A) Mouse BM monocytes were cultured in the presence of CSF1 and indicated concentrations of human ac-LDL, prior to analyzed for expression of indicated genes by qPCR.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 2 experiments.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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One-way ANOVA with Bonferroni post-test compared with 0 ng/mL. (B) NicheNet circos plot highlighting conserved ligand-receptor pairs and induced target genes between KCs and indicated niche cells in human and mouse. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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C) Feature plots showing expression of ALK1 (Acvrl1) in human myeloid cells (left) and GDF2/BMP10 in CD45 cells (right). (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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D) Livers were harvested from Fcgr1-CrexAcvrl1 mice or Acvrl1 or Acvrl1 controls and KCs examined (left) and quantified (right) using VSIG4 expression. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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E) Expression of indicated KC markers by mac populations in Fcgr1-CrexAcvrl1 or Acvrl control mice.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 3 independent experiments with n = 9 per group.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Student’s t test. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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F) Expression of indicated markers in livers of Fcgr1-CrexAcvrl1 or Acvrl1 or Acvrl1 control mice by confocal microscopy.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Scale bars, 50 μm.
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PMC8809252
|
Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
|
Images are representative of 2 mice per group. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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G) Schematic of chimera experiment setup. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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H) % chimerism normalized to levels in blood Ly6C monocytes in Clec4f-Dtr mice 7 or 13 days after DT administration following partial irradiation and receiving either Acvrl or Fcgr1-CrexAcvrl BM 4 weeks earlier.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Data are pooled from 2 independent experiments with n = 10–12 mice per group. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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I) Schematic of Fc trap experiment setup. (
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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J) Representative FACS plots showing VSIG4 and CLEC2 expression by total macs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Numbers represent % of total mac population in the indicated gate (left) and % of VSIG4 and VSIG4 macs among total CD45 cells in the different treatment conditions.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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One-way ANOVA with Bonferroni post-test.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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p < 0.05, p < 0.01, p < 0.001, p < 0.0001.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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See also Figure S8.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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To generate a practical cellular atlas of any human tissue and unravel the cell-cell circuits essential for the identities of cells inhabiting that tissue, four key pieces of information are required: (1) an inventory of all cells present, (2) the location of the different cells within the tissue to identify interactions between neighboring cells, (3) an alignment between the human and animal models allowing for any predicted cell-cell interactions to be perturbed, and (4) the identification of reliable antibody-based panels for the efficient screening of different patients and/or transgenic animals.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Here, by integrating single-cell and spatial transcriptomic and proteomic data, we provide these 4 pieces of information for the liver and uncover evolutionarily conserved microenvironmental circuits controlling the development of hepatic macs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Unraveling the spatial localization of all hepatic cells, we identify LAMs around the bile ducts in the healthy mouse, human, and macaque liver.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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However, when steatosis is present, LAMs are preferentially recruited to the steatotic regions of the liver.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This spatial information at least partially invalidates the hypothesis that LAM identity is specifically induced by fibrotic SCs (Ramachandran et al., 2019).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Rather, our data suggest that LAMs are induced by local lipid exposure.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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We also provide an alignment of the liver atlas across seven species.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This reveals the conserved and unique transcriptomic programs of steady-state KCs and uncovers the spatially restricted and conserved ligand-receptor pairs between KCs and the cells constituting their niche.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Underlining the need to first characterize the healthy tissue before attempting to understand how disease perturbs the cells, we identify the DLL-NOTCH interaction to be an evolutionarily conserved cross-talk between homeostatic LSECs and KCs and therefore not unique to hepatocellular carcinoma or fibrosis, as proposed (Ramachandran et al., 2019; Sharma et al., 2020).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Similarly, we find that FOLR2 expression is not specific to tumor-associated hepatic macs (Sharma et al., 2020) but is expressed by KCs in the healthy mouse and human liver.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Finally, we apply a proteogenomic pipeline starting from broad oligo-conjugated antibody panels for both single-cell and spatial profiling.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This is crucial as transcriptomic profiling does not always correspond with the ability to detect proteins by flow cytometry or microscopy.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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By screening broadly, we identify the best surface markers for the isolation and localization of hepatic macs and their respective niche cells.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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This allows both the validation of the spatial location at the single-cell level, and the efficient screening of transgenic mouse models for the loss of KCs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Characterization of Fcgr1-CrexAcvrl1 mice using our defined panel readily demonstrates the cruciality of the ALK1-BMP9/10 axis in KC development emphasizing that mac-stromal-cell cross-talk goes much further than the exchange of growth factors (Guilliams et al., 2020; Zhou et al., 2018).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Moving forward, applying these relatively cheap antibody panels to large patient cohorts or multiple transgenic mouse models should enable any perturbations disturbing liver homeostasis to be efficiently identified.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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The current study has two main limitations.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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First, the analysis of the human liver remains restricted by the number of human patients included (19 patients for the sc/snRNA-seq, 4 patients for Visium, and 15 patients by microscopy).
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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While this study provides markers to cheaply and efficiently screen large patient cohorts allowing this analysis to be extended, given the heterogeneity between patients, multiple studies will need to be integrated in a single analysis if we are to be able to interrogate transcriptomic differences in a particular cell subset related to age, sex, ethnicity, or pathological parameters.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Second, this study highlights that more research will be needed to fully characterize human SCs.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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As we could only retrieve these cells through snRNA-seq this means we have not been able to identify good surface markers through CITE-seq.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Additionally, for small populations of fibroblasts we did not recover enough nuclei to truly probe their heterogeneity.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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Better isolation protocols are required to retrieve and enrich these cells from the human liver in order to run a broad panel of CITE-seq markers to identify the different subsets and their corresponding surface markers.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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REAGENT or RESOURCESOURCEIDENTIFIERAntibodies:Flowcytometry/confocalmicroscopyRat Monoclonal CD102-FITC (3C4)Biolegend105606; RRID: AB_313199Rat Monoclonal CD103-BV750 (M290)BD Biosciences747478; RRID: AB_2872154Rat Monoclonal CD11b-BUV395 (M1/70)BD Biosciences565976; RRID: AB_2721166Rat Monoclonal CD11b-BV605 (M1/70)BD Biosciences563015; RRID: AB_2737951Rat Monoclonal MHCII-BUV805 (M5/114,15,2)BD Biosciences748844; RRID: AB_2873247Rat Monoclonal MHCII-FITC (M5/114,15,2)Thermo Fisher Scientific11-5321-85; RRID: AB_465233Rat Monoclonal CD14-PE Dazzle 594 (Sa14-2)Biolegend123325; RRID: AB_2721697Rat Monoclonal CD43-BUV737 (S7)BD Biosciences612840; RRID: AB_2870162Rat Monoclonal Ly6C-eFluor450 (HK1.4)eBioscience48-5932-82; RRID: AB_10805519Armenian Hamster Monoclonal CD11c-PE-Cy7 (N418)eBioscience25-0114-82; RRID: AB_469590Armenian Hamster Monoclonal CD11c-Unconjugated (N418)Biolegend117302; RRID: AB_313771Rat Monoclonal CD86-BV605 (GL-1)Biolegend105037; RRID: AB_11204429Rat Monoclonal CD54-PE (YN1/1.7.4)Biolegend116107; RRID: AB_313698Rat Monoclonal CD54-AF488 (YN1/1,7,4)Biolegend116111; RRID: AB_493494Rat Monoclonal CD172a-BB630P (P84)BD Customs624294Mouse Monoclonal XCR1 - BV650 (ZET)BioLegend148220; RRID: AB_2566410Rat Monoclonal Tim4-PerCP-Cy5.5 (RMT4-54)eBioscience46-5866-82; RRID: AB_2573781Rat Monoclonal VSIG4-PECy7 (NLA14)eBioscience25-5752-82; RRID: AB_2637431Rat Monoclonal CD19-PECy5 (1D3)eBioscience15-0193-82; RRID: AB_657672Rat Monoclonal CD19-Unconjugated (1D3)BD Biosciences553783; RRID: AB_395047Rat Monoclonal CD26-FITC (H194-112)BD Biosciences559652; RRID: AB_398295Rat Monoclonal CD38-AF700 (90)eBioscience56-0381-82; RRID: AB_657740Armenian Hamster Monoclonal CD3e-PECy5 (145-2C11)TONBO Biosciences55-0031; RRID: AB_2621815Rat Monoclonal CD3-Unconjugated (17A2)BiocerosN/ARat Monoclonal CD45-BV510 (30-F11)BioLegend103138; RRID: AB_2563061Mouse Monoclonal CD45.1-PE (A20)BD Biosciences553776; RRID: AB_395044Mouse Monoclonal CD45.1-Unconjugated (A20)Biolegend110702; RRID: AB_313491Mouse Monoclonal CD45.2-AF700 (104)eBioscience56-0454-82; RRID: AB_657752Rat Monoclonal CD45R-PE-Cy5 (RA3-6B2)BD Biosciences553091; RRID: AB_394621Rat Monoclonal Ly6G-BUV563 (1A8)BD Biosciences612921; RRID: AB_2870206Mouse Monoclonal CD64-BV711 (X54-5/7.1)BioLegend139311; RRID: AB_2563846Mouse Monoclonal NK1.1-PECy5 (PK136)BioLegend108716; RRID: AB_493590Rat Monoclonal Ter119-PECy5 (TER-119)eBioscience15-5921-82; RRID: AB_468810Rat Monoclonal F4/80-BV785 (BM8)BioLegend123141; RRID: AB_2563667Rat Monoclonal F4/80-Unconjugated (CI:A3-1)Bio-RadMCA497R; RRID: AB_323279Rat Monoclonal F4/80-AF594 (BM8)Biolegend123140; RRID: AB_2563241Rat Monoclonal CLEC2-PE (17D9)Biolegend146104; RRID: AB_2562382Rat Monoclonal FOLRB-PE (10/FR2)Biolegend153304; RRID: AB_2721344Rat Monoclonal FOLRB-Unconjugated (10/FR2)Biolegend153302; RRID: AB_2687271Rat Monoclonal SiglecF-BUV395 (E50-2440)BD Biosciences740280; RRID: AB_2740019Rat Monoclonal CD207-AF647ImgenexDDX0362A647; RRID: AB_1148741Rat Monoclonal CD206-AF647Biolegend141712; RRID: AB_10900420Rat Monoclonal CD326 (EPCAM)-APC (G8.8)eBioscience17-5791-82; RRID: AB_2716944Goat Polyclonal Clec4F-UnconjugatedR & D SystemsAF2784; RRID: AB_2081339Rat Monoclonal Clec4F-Unconjugated (370901)R & D SystemsMAB2784; RRID: AB_2081338Goat Polyclonal VSIG4-UnconjugatedR & D SystemsAF4646; RRID: AB_2257239Goat Polyclonal Lyve1-UnconjugatedR&D SystemsAF2125; RRID: AB_2297188Donkey Anti-Goat IgG-AF488Thermo Fisher ScientificA-11055; RRID: AB_2534102Donkey Anti-Goat IgG–AF633Thermo Fisher ScientificA-21082; RRID: AB_2535739Donkey Anti-Goat IgG-AF647Thermo Fisher ScientificA-21447; RRID: AB_2535864Donkey Anti-Rat IgG-Cy3Jackson ImmunoResearch712-166-153; RRID: AB_2340669Donkey Anti-Mouse IgG–AF555Thermo Fisher ScientificA-31570; RRID: AB_2536180Donkey Anti-Rabbit IgG-AF647Thermo Fisher ScientificA-31573; RRID: AB_2536183Donkey Anti-Rabbit IgG-AF680Thermo Fisher ScientificA-32802; RRID: AB_2762836Goat Anti-Rabbit IgG–AF514Thermo Fisher ScientificA-31558; RRID: AB_2536173Goat Anti-Hamster IgG-Cy3Jackson ImmunoResearch127-165-160; RRID: AB_2338989Goat Anti-Hamster IgG-AF594Jackson ImmunoResearch127-585-160; RRID: AB_2338999Goat Anti-Rat IgG-AF568Thermo Fisher ScientificA-11077; RRID: AB_2534121Goat Anti-Chicken IgY-AF680AbcamAb175779;Rat Monoclonal CD31-Unconjugated (MEC13.3)BD Biosciences550274; RRID: AB_393571Rabbit Polyclonal Desmin-UnconjugatedAbcamab15200; RRID: AB_301744Rabbit Polyclonal Glutamine Synthetase - UnconjugatedAbcamab73593; RRID: AB-2247588Goat Anti-Rabbit IgG-AF514Thermo Fisher ScientificA-31558; RRID: AB_2536173Rabbit Monoclonal GPNMB- unconjugated (EPR18226-147)Abcamab188222;Polyclonal Chicken anti-GFPAves LabsGFP-1010; RRID: AB_2307313Mouse Monoclonal FOLR2-PE (94b/FOLR2)Biolegend391704; RRID: AB_2721336Mouse Monoclonal CD45-APCCy7 (2D1)Biolegend368516; RRID: AB_2566376Mouse Monoclonal CD14-PETexasRed (TuK4)Thermo Fisher ScientificMHCD1417; RRID:AB_10373552Mouse Monoclonal CD1C-Unconjugated (L161)Biolegend331502; RRID:AB_1088995Mouse Monoclonal CD163-Unconjugated (10D6)Thermo Fisher ScientificMA5-11458; RRID: AB_10982556Mouse Monoclonal EPCAM-eF660 (1B7)eBioscience50-9326-42: RRID: AB_10598658Rabbit Monoclonal CD68-Unconjugated (EPR20545)AbcamAb213363; RRID: AB_2801637Rabbit Monoclonal CD34-PE (EP373Y)AbcamAb223930;Anti- Mouse CD38 (REA616) FITCMiltenyi Biotec130-122-955; RRID: AB_2811415Anti-Mouse CD26 (REA1196) PEMiltenyi Biotec130-122-775; RRID: AB_2801934Anti-Mouse F4/80 (REA126) FITCMiltenyi Biotec130-117-509; RRID: AB_2727970Anti-Mouse CD5 (53-7.3) FITCMiltenyi Biotec130-102-574; RRID: AB_2658608Anti-Mouse CD79B (REA1117) PEMiltenyi Biotec130-119-425; RRID: AB_2751702Anti-Mouse CD45R/B220 (RA3-6B2) FITCMiltenyi Biotec130-118-323; RRID: AB_2751482Anti-Mouse CD43 (L11) PEMiltenyi Biotec130-102-594; RRID: AB_2661309Anti-Mouse CCR2 (REA538) PEMiltenyi Biotec130-117-548; RRID: AB_2727981Anti-Mouse MHCII (M5/114.15.2) FITCMiltenyi Biotec130-123-666; RRID: AB_2802055Anti-Mouse CD73 (REA778) PEMiltenyi Biotec130-111-331; RRID: AB_2659153Anti-Mouse CD146 (REA1064) FITCMiltenyi Biotec130-118-252; RRID: AB_2751472Anti-Mouse CD90.2 (30-H12) PEMiltenyi Biotec130-120-091; RRID: AB_2751997Anti-Mouse CD138 ( REA104) PEMiltenyi Biotec130-120-810; RRID: AB_2752204Anti-Mouse CD29 (REA1074) PEMiltenyi Biotec130-119-165; RRID: AB_2751649Anti-Mouse CD105 (REA1058) FITCMiltenyi Biotec130-118-173; RRID: AB_2733613Anti-Human CD36 (REA760) PEMiltenyi Biotec130-110-877; RRID: AB_2657728Anti-Human CD206 (REAL518) APCMiltenyi Biotec130-122-168; RRID: AB_2857557Anti-Human Collagen IV (REAL567) PEMiltenyi Biotec130-122-866; RRID: AB_2857566Anti-Human CD105 (REA794) PEMiltenyi Biotec130-112-163; RRID: AB_2654424Anti-Human CD146 (REA773) APCMiltenyi Biotec130-111-323; RRID: AB_2655179Anti-Human CD90 (REA897) PEMiltenyi Biotec130-114-860; RRID: AB_2726811Anti-Human DESMIN (REA1134) PEMiltenyi Biotec130-119-490; RRID: AB_2857461Anti-Human CD74 (REA1103) PEMiltenyi Biotec130-119-203; RRID: AB_2733836Anti-Human EPCAM (REA764) FITCMiltenyi Biotec130-110-998; RRID: AB_2657493Anti-Human CD68 (REAL566) PEMiltenyi Biotec130-123-368; RRID: AB_2857592Anti-Human CD1c (REA694) APCMiltenyi Biotec130-110-537; RRID: AB_2656040Anti-Human CD3 (REA1151) APCMiltenyi Biotec130-120-269; RRID: AB_2876933Anti-Human CD5 (REA782) PEMiltenyi Biotec130-110-990; RRID: AB_2658593Anti-Human CD7 (REA1244) PEMiltenyi Biotec130-124-939; RRID: AB_2819716Anti-Human CD19 (REAL106) PEMiltenyi Biotec130-122-649; RRID: AB_2784034Anti-Human CD22 (REA340) APCMiltenyi Biotec130-120-762; RRID: AB_2752186Anti-Human S100A8 (REA917) PEMiltenyi Biotec130-115-253; RRID: AB_2726963Anti-Human CD177 (REA258) PEMiltenyi Biotec130-101-525; RRID: AB_2655641Anti-Human CD169 (REA1176) PEMiltenyi Biotec130-121-115; RRID: AB_2783985Anti-Human CD163 (REA406) PEMiltenyi Biotec130-121-316; RRID: AB_2857545Antibodies:CITE-seqTotalSeq-A0001 anti-mouse CD4 (RM4-5)BioLegend100569; RRID: AB_2749956TotalSeq-A0002 anti-mouse CD8a (53-6.7)BioLegend100773; RRID: AB_2734151TotalSeq-A0003 anti-mouse CD366 (Tim-3) (RMT3-23)BioLegend119729; RRID: AB_2734178TotalSeq-A0004 anti-mouse CD279 (PD-1) (RMP1-30)BioLegend109123; RRID: AB_2734169TotalSeq-A0005 anti-human CD80 (2D10)BioLegend305239; RRID: AB_2749958TotalSeq-A0006 anti-human CD86 (IT2.2)BioLegend305443; RRID: AB_2734273TotalSeq-A0007 anti-human CD274 (29E.2A3)BioLegend329743; RRID: AB_2749959TotalSeq-A0008 anti-human CD273 (24F.10C112)BioLegend329619; RRID: AB_2734321TotalSeq-A0009 anti-human CD275 (2D3)BioLegend309413; RRID: AB_2734278TotalSeq-A0010 anti-human CD276 (DCN.70)BioLegend331607; RRID: AB_2734327TotalSeq-A0012 anti-mouse CD117 (c-kit) (2B8)BioLegend105843; RRID: AB_2749960TotalSeq-A0013 anti-mouse Ly-6C (HK1.4)BioLegend128047; RRID: AB_2749961TotalSeq-A0014 anti-mouse/human CD11b (M1/70)BioLegend101265; RRID: AB_2734152TotalSeq-A0015 anti-mouse Ly-6G (1A8)BioLegend127655; RRID: AB_2749962TotalSeq-A0016 anti-human Galectin9 (9M1-3)BioLegendBarcode: ACTCACTGGAGTCTCTotalSeq-A0020 anti-human CD270 (122)BioLegend318813; RRID: AB_2734293TotalSeq-A0021 anti-human CD252 (11C3.1)BioLegendBarcode: TTTAGTGATCCGACTTotalSeq-A0022 anti-human CD137L (5F4)BioLegend311509; RRID: AB_2734284TotalSeq-A0023 anti-human CD155 (SKII.4)BioLegend337623; RRID: AB_2749963TotalSeq-A0024 anti-human CD112 (TX31)BioLegend337417; RRID: AB_2749964TotalSeq-A0026 anti-human CD47 (CC2C6)BioLegend323129; RRID: AB_2734305TotalSeq-A0027 anti-human CD70 (113-16)BioLegend355117; RRID: AB_2749965TotalSeq-A0028 anti-human CD30 (BY88)BioLegend333913; RRID: AB_2749966TotalSeq-A0029 anti-human CD48 (BJ40)BioLegend336709; RRID: AB_2734342TotalSeq-A0031 anti-human CD40 (5C3)BioLegend334346; RRID: AB_2749968TotalSeq-A0032 anti-human CD154 (24-31)BioLegend310843; RRID: AB_2734283TotalSeq-A0033 anti-human CD52 (HI186)BioLegend316017; RRID: AB_2734292TotalSeq-A0034 anti-human CD3 (UCHT1)BioLegend300475; RRID: AB_2734246TotalSeq-A0047 anti-human CD56 (5.1H11)BioLegend362557; RRID: AB_2749970TotalSeq-A0050 anti-human CD19 (HIB19)BioLegend302259; RRID: AB_2734256TotalSeq-A0052 anti-human CD33 (P67.6)BioLegend366629; RRID: AB_2734409TotalSeq-A0053 anti-human CD11c (S-HCL-3)BioLegend371519; RRID: AB_2749971TotalSeq-A0054 anti-human CD34 (581)BioLegend343537; RRID: AB_2749972TotalSeq-A0056 anti-human CD269 (19F2)BioLegend357521; RRID: AB_2749974TotalSeq-A0057 anti-human B2M (2M2)BioLegend316321; RRID: AB_2749975TotalSeq-A0058 anti-human HLA-ABC (W6/32)BioLegend311445; RRID: AB_2749976TotalSeq-A0060 anti-human CD90 (5E+10)BioLegend328135; RRID: AB_2734312TotalSeq-A0061 anti-human CD117 (104D2)BioLegend313241; RRID: AB_2734287TotalSeq-A0062 anti-human CD10 (HI10a)BioLegend312231; RRID: AB_2832627TotalSeq-A0063 anti-human CD45RA (HI100)BioLegend304157; RRID: AB_2734267TotalSeq-A0064 anti-human CD123 (6H6)BioLegend306037; RRID: AB_2749977TotalSeq-A0066 anti-human CD7 (CD7-6B7)BioLegend343123; RRID: AB_2734345TotalSeq-A0068 anti-human CD105 (43A3)BioLegend323221; RRID: AB_2750350TotalSeq-A0069 anti-human CD201 (RCR-401)BioLegend351907; RRID: AB_2749978TotalSeq-A0070 anti-human/mouse CD49f (GoH3)BioLegend313633; RRID; AB_2734291TotalSeq-A0073 anti-mouse/human CD44 (IM7)BioLegend103045; RRID: AB_2734154TotalSeq-A0074 anti-mouse CD54 (YN1/1.7.4)BioLegend116127; RRID: AB_2734177TotalSeq-A0074 anti-mouse CD90.2 (30-H12)Biolegend105345; RRID: AB_2734166TotalSeq-A0076 anti-mouse/human CD15 (SSEA-1) (MC-480)BioLegend125615; RRID: AB_2800603TotalSeq-A0077 anti-mouse CD73 (TY/11.8)BioLegend127227; RRID: AB_2749980TotalSeq-A0078 anti-mouse CD49d (R1-2)BioLegend103623; RRID: AB_2734159TotalSeq-A0079 anti-mouse CD200 (OX2) (OX-90)BioLegend123811; RRID: AB_2734191TotalSeq-A0080 anti-human CD8a (RPA-T8)BioLegend301067; RRID: AB_2734248TotalSeq-A0081 anti-human CD14 (M5E2)BioLegend301855: RRID: AB_2734254TotalSeq-A0083 anti-human CD16 (3G8)BioLegend302061; RRID: AB_2734255TotalSeq-A0085 anti-human CD25 (BC96)BioLegend302643; RRID: AB_2734258TotalSeq-A0087 anti-human CD45RO (UCHL1)BioLegend304255; RRID: AB_2734268TotalSeq-A0088 anti-human CD279 (EH12.2H7)BioLegend329955; RRID: AB_2734322TotalSeq-A0089 anti-human TIGIT (A15153G)BioLegend372725; RRID: AB_2734426TotalSeq-A0090 Mouse IgG1, κ isotype Ctrl (MOPC-21)BioLegend400199; RRID: AB_2868412TotalSeq-A0091 Mouse IgG2a, κ isotype Ctrl (MOPC-173)BioLegend400285;TotalSeq-A0092 Mouse IgG2b, κ isotype Ctrl (MPC-11)BioLegend400373;TotalSeq-A0093 anti-mouse CD19 (B4)BioLegend115559; RRID: AB_2749981TotalSeq-A0094 anti-mouse CD3e (145-2C11)BioLegendBarcode: TAATGCCAGTTGTGCTotalSeq-A0095 Rat IgG2b, κ Isotype Ctrl (RTK4530)BioLegend400673;TotalSeq-A0097 anti-mouse CD25 (PC61)BioLegend102055; RRID: AB_2749982TotalSeq-A0098 anti-mouse CD135 (A2F10)BioLegend135316; RRID: AB_2749983TotalSeq-A0100 anti-human CD20 (2H7)BioLegend302359; RRID: AB_2749984TotalSeq-A0101 anti-human CD335 (900)BioLegend331943; RRID: AB_2800875TotalSeq-A0102 anti-human CD294 (BM16)BioLegend350127; RRID: AB_2734360TotalSeq-A0103 anti-mouse/human CD45R/B220BioLegend103263; RRID: AB_2734158TotalSeq-A0104 anti-mouse CD102 (3C4 (MIC2/4))BioLegend105613; RRID: AB_2734167TotalSeq-A0105 anti-mouse CD115 (CSF-1R) (AFS98)BioLegend135533; RRID: AB_2734198TotalSeq-A0106 anti-mouse CD11c (N418)BioLegend117355; RRID: AB_2750352TotalSeq-A0107 anti-mouse CD21/CD35 (CR2/CR1) (7E9)BioLegend123427; RRID: AB_2750540TotalSeq-A0108 anti-mouse CD23 (B3B4)BioLegend101635; RRID: AB_2750358TotalSeq-A0109 anti-mouse CD16/32 (93)BioLegend101343; RRID: AB_2750532TotalSeq-A0110 anti-mouse CD43 (S11)BioLegend143211; RRID: AB_2750541TotalSeq-A0111 anti-mouse CD5 (53-7.3)BioLegend100637; RRID: AB_2749985TotalSeq-A0112 anti-mouse CD62L (MEL-14)BioLegend104451; RRID: AB_2750364TotalSeq-A0113 anti-mouse CD93 (AA4.1, early B lineage) (AA4.1)BioLegend136513; RRID: AB_2750375TotalSeq-A0114 anti-mouse F4/80 (BM8)BioLegend123153; RRID: AB_2749986TotalSeq-A0115 anti-mouse FcεRIα (MAR-1)BioLegend134333; RRID: AB_2749987TotalSeq-A0117 anti-mouse I-A/I-E (M5/114.15.2)BioLegend107653; RRID: AB_2750505TotalSeq-A0118 anti-mouse NK-1.1 (PK136)BioLegend108755; RRID: AB_2750536TotalSeq-A0119 anti-mouse Siglec H (551)BioLegend129615; RRID: AB_2750537TotalSeq-A0120 anti-mouse TCRb (H57-597)Biolegend109247; RRID: AB_2750538TotalSeq-A0121 anti-mouse TCRgd (GL3)Biolegend118137; RRID: AB_2749988TotalSeq-A0122 anti-mouse Ter-119 (Ter-119)Biolegend116247; RRID: AB_2749989TotalSeq-A0123 anti-human CD326 (9C4)BioLegend324241; RRID: AB_2750362TotalSeq-A0124 anti-human CD31 (WM59)BioLegend303137: RRID: AB_2750360TotalSeq-A0126 anti-human CD133 (clone 7)BioLegend372815; RRID: AB_2750353TotalSeq-A0127 anti-human Podoplanin (NC-08)BioLegend337019; RRID; AB_2749990TotalSeq-A0128 anti-human CD140a (16A1)BioLegend323509; RRID: AB_2750354TotalSeq-A0129 anti-human CD140b (18A2)BioLegend323609; RRID: AB_2783192TotalSeq-A0130 anti-mouse Ly-6A/E (Sca-1) (D7)BioLegend108147; RRID: AB_2750535TotalSeq-A0131 anti-human Cadherin (16G5)BioLegend368715; RRID: AB_2749991TotalSeq-A0132 anti-human EGFR (AY13)BioLegend352923; RRID: AB_2734373TotalSeq-A0134 anti-human CD146 (P1H12)BioLegend361017: RRID: AB_2750355TotalSeq-A0136 anti-human IgM (MHM-88)BioLegend314541; RRID: AB_2749992TotalSeq-A0138 anti-human CD5 (UCHT2)BioLegend300635; RRID: AB_2750346TotalSeq-A0139 anti-human TCR (B1)BioLegend331229; RRID: AB_2734325TotalSeq-A0140 anti-human CD183 (G025H7)BioLegend353745; RRID: AB_2749993TotalSeq-A0141 anti-human CD195 (J418F1)BioLegend359135; RRID: AB_2749994TotalSeq-A0142 anti-human CD32 (FUN-2)BioLegend303223; RRID: AB_2749995TotalSeq-A0143 anti-human CD196 (G034E3)BioLegend353437; RRID: AB_2750534TotalSeq-A0144 anti-human CD185 (J252D4)BioLegend356937; RRID: AB_2750356TotalSeq-A0145 anti-human CD103 (Ber-ACT8)BioLegend350231; RRID: AB_2749996TotalSeq-A0146 anti-human CD69 (FN50)BioLegend310947; RRID: AB_2749997TotalSeq-A0147 anti-human CD62L (DREG-56)BioLegend304847; RRID: AB_2750365TotalSeq-A0148 anti-human CD197 (G043H7)BioLegend353247; RRID: AB_2750357TotalSeq-A0149 anti-human CD161 (HP-3G10)BioLegend339945; RRID: AB_2749998TotalSeq-A0151 anti-human CD152 (BNI3)BioLegend369619; RRID: AB_2734423TotalSeq-A0152 anti-human CD223 (11C3C65)BioLegend369333; RRID: AB_2749999TotalSeq-A0153 anti-human KLRG1 (SA231A2)BioLegend367721; RRID: AB_2750373TotalSeq-A0155 anti-human CD107a (H4A3)BioLegend328647; RRID: AB_2750351TotalSeq-A0156 anti-human CD95 (DX2)BioLegend305649; RRID: AB_2750368TotalSeq-A0158 anti-human CD134 (Ber-ACT35)BioLegend350033; RRID: AB_2783245TotalSeq-A0159 anti-human HLA-DR (L243)BioLegend307659; RRID: AB_2750001TotalSeq-A0160 anti-human CD1c (L161)BioLegend331539; RRID: AB_2734326TotalSeq-A0162 anti-human CD64 (10.1)BioLegend305037; RRID: AB_2750366TotalSeq-A0163 anti-human CD141 (M80)BioLegend344121; RRID: AB_2783229TotalSeq-A0164 anti-human CD1d (51.1)BioLegend350317; RRID: AB_2750370TotalSeq-A0165 anti-human CD314 (1D11)BioLegend320835; RRID: AB_2734298TotalSeq-A0166 anti-human CD66b (6/40c)BioLegend392905; RRID: AB_2750372TotalSeq-A0167 anti-human CD35 (E11)BioLegend333407; RRID: AB_2783217TotalSeq-A0168 anti-human CD57 (QA17A04)BioLegend393319; RRID: AB_2810588TotalSeq-A0169 anti-human CD366 (F38-2E2)BioLegend345047; RRID: AB_2800924TotalSeq-A0170 anti-human CD272 (MIH26)BioLegend344525; RRID: AB_2750002TotalSeq-A0171 anti-human/mouse/rat CD278 (ICOS) (C398.4A)BioLegend313555; RRID: AB_2800824TotalSeq-A0173 anti-mouse CD206 (MMR) (C068C2)BioLegendBarcode: TCAACTCGGTGTTGCTotalSeq-A0174 anti-human CD58 (TS2/9)BioLegend330919; RRID: AB_2750003TotalSeq-A0175 anti-human CD96 (NK92.39)BioLegend338419; RRID: AB_2750004TotalSeq-A0176 anti-human CD39 (A1)BioLegend328233; RRID: AB_2750005TotalSeq-A0177 anti-human CD178 (NOK-1)BioLegend306413; RRID: AB_2750500TotalSeq-A0180 anti-human CD24 (ML5)BioLegend311137: RRID: AB_2750374TotalSeq-A0181 anti-human CD21 (Bu32)BioLegend354915; RRID: AB_2750006TotalSeq-A0182 anti-mouse CD3 (17A2)Biolegend100251; RRID: AB_2750533TotalSeq-A0184 anti-mouse CD335 (NKp46) (29A1.4)BioLegend137633; RRID: AB_2734199TotalSeq-A0185 anti-human CD11a (TS2/4)BioLegend350615; RRID: AB_2734365TotalSeq-A0186 anti-human IgA (HP6123)BioLegendBarcode: AAGATGTCCGAGCAATotalSeq-A0187 anti-human CD79b (CB3-1)BioLegend341415; RRID: AB_2750347TotalSeq-A0188 anti-human CD66a_c_e (ASL-32)BioLegend342319; RRID: AB_2783223TotalSeq-A0189 anti-human CD244 (C1.7)BioLegend329527; RRID: AB_2750007TotalSeq-A0190 anti-mouse CD274 (B7-H1, PD-L1) (MIH6)BioLegend153604; RRID: AB_2783125TotalSeq-A0191 anti-mouse/rat/human CD27 (LG.3A10 )BioLegend124235; RRID: AB_2750344TotalSeq-A0192 anti-mouse CD20 (SA275A11)BioLegend150423; RRID: AB_2734214TotalSeq-A0193 anti-mouse CD357 (GITR) (DTA-1)BioLegend126319; RRID: AB_2734195TotalSeq-A0194 anti-mouse CD137 (17B5)BioLegend106111; RRID: AB_2783048TotalSeq-A0195 anti-mouse CD134 (OX-40) (OX-86)BioLegend119426; RRID: AB_2750376TotalSeq-A0196 anti-human CD235ab (HIR2)BioLegend306623; RRID: AB_2750008TotalSeq-A0197 anti-mouse CD69 (H1.2F3)BioLegend104546; RRID: AB_2750539TotalSeq-A0198 anti-mouse CD127 (IL-7Rα) (A7R34)BioLegend135045; RRID: AB_2750009TotalSeq-A0200 anti-mouse CD86 (GL-1)BioLegend105047; RRID: AB_2750348TotalSeq-A0201 anti-mouse CD103 (2E7)BioLegend121437; RRID: AB_2750349TotalSeq-A0202 anti-mouse CD64 (FcγRI) (X54-5/7.1)BioLegend139325; RRID: AB_2750367TotalSeq-A0203 anti-mouse CD150 (SLAM) (TC15-12F12.2)BioLegend115945; RRID: AB_2783055TotalSeq-A0204 anti-mouse CD28 (37.51)BioLegendBarcode: ATTAAGAGCGTGTTGTotalSeq-A0205 anti-human CD206 (15-2)BioLegend321143; RRID: AB_2750010TotalSeq-A0206 anti-human CD169 (7-239)BioLegend346011; RRID: AB_2750011TotalSeq-A0207 anti-human CD370 (8F9)BioLegend353807; RRID: AB_2814293TotalSeq-A0208 anti-human XCR1 (S15046E)BioLegend372613; RRID: AB_2783286TotalSeq-A0209 anti-mouse TCR (2.11)BioLegend141113; RRID: AB_2800654TotalSeq-A0210 anti-mouse TCR (536)BioLegend137507; RRID: AB_2810404TotalSeq-A0211 anti-mouse TCR (UC3-10A6)BioLegend137709; RRID: AB_2783100TotalSeq-A0212 anti-mouse CD24 (M1/69)BioLegend101841; RRID: AB_2750380TotalSeq-A0213 anti-human Notch (MHN1-519)BioLegend352109; RRID: AB_2783247TotalSeq-A0214 anti-human/mouse integrin β7 (FIB504)BioLegend321227; RRID: AB_2750504TotalSeq-A0215 anti-human CD268 (11C1)BioLegend316925; RRID: AB_2750502TotalSeq-A0216 anti-human CD42b (HIP1)BioLegend303937; RRID: AB_2783163TotalSeq-A0217 anti-human CD54 (HA58)BioLegend353123; RRID: AB_2750384TotalSeq-A0218 anti-human CD62P (AK4)BioLegend304933; RRID: AB_2750386TotalSeq-A0219 anti-human CD119 (GIR-208)BioLegend308607; RRID: AB_2750385TotalSeq-A0221 anti-human IRF5 (11F4A09)BioLegendBarcode: GGTAGCCCTAGAGTATotalSeq-A0224 anti-human TCR (IP26)BioLegend306737: RRID: AB_2783167TotalSeq-A0225 anti-mouse CD196 (CCR6) (29-2L17)BioLegend129825; RRID: AB_2783083TotalSeq-A0226 anti-mouse CD106 (429 (MVCAM.A))BioLegend105725; RRID: AB_2783044TotalSeq-A0227 anti-mouse CD122 (IL-2Rb) (5H4)BioLegendBarcode: GGTATGCGACACTTATotalSeq-A0228 anti-mouse CD183 (CXCR3) (CXCR3-173)BioLegendBarcode: GTTCACGCCGTAGACTTotalSeq-A0229 anti-mouse CD62P (P-selectin) (RMP-1)BioLegendBarcode: TGTGTGCCGTAGACTTotalSeq-A0230 anti-mouse CD8b (Ly-3) (YTS156.7.7)BioLegend126623; RRID: AB_2800615TotalSeq-A0232 anti-mouse MAdCAM-1 (MECA-367)BioLegend120713; RRID: AB_2783058TotalSeq-A0235 anti-mouse TCR (KJ16-133.18)BioLegend118415; RRID: AB_2783056TotalSeq-A0236 Rat IgG1, κ Isotype Ctrl (RTK2071)BioLegend400459;TotalSeq-A0237 Rat IgG1, λ Isotype Ctrl (G0114F7)BioLegend401919;TotalSeq-A0238 Rat IgG2a, κ Isotype Ctrl (RTK2758)BioLegend400571;TotalSeq-A0240 Purified Rat IgG2c, κ Isotype Ctrl (RTK4174)BioLegend400739;TotalSeq-A0241 Armenian Hamster IgG Isotype Ctrl (HTK888)BioLegend400973;TotalSeq-A0242 anti-human CD192 (K036C2)BioLegend357229; RRID: AB_2750501TotalSeq-A0244 anti-human CD102 (CBR-IC2/2)BioLegend328509; RRID: AB_2750381TotalSeq-A0245 anti-human CD106 (STA)BioLegend305813; RRID: AB_2800788TotalSeq-A0247 anti-human CD267 (1A1)BioLegend311913; RRID: AB_2783181TotalSeq-A0248 anti-human CD62E (HAE-1f)BioLegend336017; RRID: AB_2783218TotalSeq-A0249 anti-mouse/human IRF4 (IRF4.3E4)BioLegendBarcode: GGATTTGTATCTCCCTotalSeq-A0250 anti-mouse/human KLRG1 (MAFA) (2F1/KLRG1)BioLegend138431; RRID: AB_2800648TotalSeq-A0351 anti-human CD135 (BV10A4H2)BioLegend313317; RRID: AB_2783188TotalSeq-A0352 anti-human FCeR1a (AER-37)BioLegend334641; RRID: AB_2750503TotalSeq-A0353 anti-human CD41 (HIP8)BioLegend303737; RRID: AB_2783162TotalSeq-A0354 anti-mouse TCR (MR9-4)BioLegend139517; RRID: AB_2810408TotalSeq-A0355 anti-human CD137 (4B4-1)BioLegend309835; RRID: AB_2783173TotalSeq-A0356 anti-human CD254 (MIH24)BioLegend347509; RRID: AB_2750377TotalSeq-A0357 anti-human CD43 (CD43-10G7)BioLegend343209; RRID: AB_2800915TotalSeq-A0358 anti-human CD163 (GHI/61)BioLegend333635; RRID: AB_2750343TotalSeq-A0359 anti-human CD83 (HB15e)BioLegend305339; RRID: AB_2800784TotalSeq-A0360 anti-human CD357 (108-17)BioLegend371225; RRID: AB_2801016TotalSeq-A0361 anti-human CD59 (p282 (H19))BioLegend304709; RRID: AB_2750371TotalSeq-A0362 anti-human CD309 (7D4-6)BioLegend359919; RRID: AB_2810571TotalSeq-A0364 anti-human CD13 (WM15)BioLegend301729; RRID: AB_2783151TotalSeq-A0366 anti-human CD184 (12G5)BioLegend306531; RRID: AB_2800790TotalSeq-A0367 anti-human CD2 (TS1/8)BioLegendBarcode: TACGATTTGTCAGGGTotalSeq-A0368 anti-human CD226 (11A8)BioLegend338335; RRID: AB_2783220TotalSeq-A0369 anti-human CD29 (TS2/16)BioLegend303027; RRID: AB_2783160TotalSeq-A0370 anti-human CD303 (201A)BioLegend354239; RRID: AB_2800952TotalSeq-A0371 anti-human CD49b (P1E6-C5)BioLegend359311; RRID: AB_2750363TotalSeq-A0373 anti-human CD81 (5A6)BioLegend349521; RRID: AB_2800930TotalSeq-A0374 anti-human CD98 (MEM-108)BioLegend315605; RRID: AB_2750369TotalSeq-A0375 anti-human IgG (M1310G05)BioLegend410725; RRID: AB_2783329TotalSeq-A0376 anti-mouse CD195 (CCR5) (HM-CCR5)BioLegend107019; RRID: AB_2783049TotalSeq-A0377 anti-mouse CD197 (CCR7) (4B12)BioLegendBarcode: TTATTAACAGCCCACTotalSeq-A0378 anti-mouse CD223 (LAG-3) (C9B7W)BioLegend125229; RRID: AB_2783078TotalSeq-A0379 anti-mouse CD62E (E-selectin) (RME-1/CD62E)BioLegendBarcode: CTCCCTTTGTAACATTotalSeq-A0381 anti-mouse Panendothelial Cell Antigen (MECA-32)BioLegend120507; RRID: AB_2783057TotalSeq-A0382 anti-human CD177 (MEM-166)BioLegend315811: RRID: AB_2750554TotalSeq-A0383 anti-human CD55 (JS11)BioLegend311317; RRID: AB_2750378TotalSeq-A0384 anti-human IgD (IA6-2)BioLegend348243; RRID: AB_2783238TotalSeq-A0385 anti-human CD18 (TS1/18)BioLegend302121; RRID: AB_2750382TotalSeq-A0386 anti-human CD28 (CD28.2)BioLegend302955; RRID: AB_2783159TotalSeq-A0387 anti-human TSLPR (1D3)BioLegend322907; RRID: AB_2800845TotalSeq-A0388 anti-mouse CD152 (UC10-4B9)BioLegend106325; RRID: AB_2876417TotalSeq-A0389 anti-human CD38 (HIT2)BioLegend303541; RRID: AB_2783161TotalSeq-A0390 anti-human CD127 (A019D5)BioLegend351352; RRID: AB_2734366TotalSeq-A0392 anti-human CD15 (W6D3)BioLegend323046; RRID: AB_2734304TotalSeq-A0393 anti-human CD22 (S-HCL-1)BioLegend363514; RRID: AB_2734404TotalSeq-A0394 anti-human CD71 (CY1G4)BioLegend334123; RRID: AB_2800884TotalSeq-A0395 anti-human B7H4 (MIH43)BioLegend358114; RRID: AB_2734386TotalSeq-A0396 anti-human CD26 (BA5b)BioLegend302720; RRID: AB_2734261TotalSeq-A0397 anti-human CD193 (5e7)BioLegend310729; RRID: AB_2783174TotalSeq-A0398 anti-human CD115 (9-4D2-1E4)BioLegend347325; RRID: AB_2783237TotalSeq-A0399 anti-human CD204 (7C9C20)BioLegend371909; RRID: AB_2810584TotalSeq-A0400 anti-human CD144 (BV9)BioLegend348517; RRID: AB_2783239TotalSeq-A0401 anti-human CD301 (H037G3)BioLegend354707; RRID: AB_2810566TotalSeq-A0402 anti-human CD1a (HI149)BioLegend300133; RRID: AB_2783146TotalSeq-A0404 anti-human CD63 (H5C6)BioLegend353035; RRID: AB_2783249TotalSeq-A0405 anti-human CD284 (HTA125)BioLegend312817; RRID: AB_2783183TotalSeq-A0406 anti-human CD304 (12C2)BioLegend354525; RRID: AB_2783261TotalSeq-A0407 anti-human CD36 (5-271)BioLegend336225; RRID: AB_2800892TotalSeq-A0408 anti-human CD172a (15-414)BioLegend372109; RRID: AB_2783285TotalSeq-A0409 anti-human CD85g (17G10.2BioLegend326411; RRID: AB_2750555TotalSeq-A0411 Polyclonal Clec4F (Custom)BioLegendBarcode: TATGCTGTGGCTATGTotalSeq-A0414 Polyclonal VSIG4 (Custom)BioLegendBarcode: ACGTATATCACGTTGTotalSeq-A0415 anti-P2RY12 (S16007D)BioLegend848009; RRID: AB_2783419TotalSeq-A0416 anti-mouse CD300LG (Nepmucin) (ZAQ5)BioLegend147105; RRID: AB_2783116TotalSeq-A0417 anti-mouse CD163 (S16007D)BioLegend155303; RRID: AB_2814058TotalSeq-A0418 anti-human CD243 (4E3.16)BioLegend919407: RRID: AB_2810817TotalSeq-A0419 anti-human CD72 (3F3)BioLegend316205; RRID: AB_2783189TotalSeq-A0420 anti-human CD158 (HP-MA4)BioLegend339515; RRID: AB_2800901TotalSeq-A0421 anti-mouse CD49b (HMa2)BioLegend103523; RRID: AB_2819796TotalSeq-A0422 anti-mouse CD172a (SIRPα) (P84)BioLegend144033; RRID: AB_2800670TotalSeq-A0423 anti-human MerTK (5900H11G1E3)BioLegend367617; RRID: AB_2801011TotalSeq-A0424 anti-mouse CD14 (Sa14-2)BioLegend123333; RRID: AB_2800591TotalSeq-A0426 anti-mouse CD192 (CCR2) (SA203G11)BioLegend150625; RRID: AB_2783122TotalSeq-A0427 anti-human FOLR2 (94b/FOLR2)BioLegend391707; RRID: AB_2783289TotalSeq-A0428 anti-human TIM4 (9F4)BioLegend354009; RRID: AB_2783258TotalSeq-A0429 anti-mouse CD48 (HM48.1)BioLegend103447; RRID: AB_2800558TotalSeq-A0430 anti-human CD171 (L1-OV198.5)BioLegend371609; RRID: AB_2801019TotalSeq-A0432 anti-human CD320 (3F4)BioLegend800319; RRID: AB_2801138TotalSeq-A0433 anti-human CD325 (8C11)BioLegend350817; RRID: AB_2810555TotalSeq-A0434 anti-mouse/human ReceptorD4BioLegendBarcode: TCTCTGCACCGCTTTTotalSeq-A0435 anti-mouse/human GABRB3BioLegendBarcode: GGTGTGAGCAGTTCTTotalSeq-A0437 anti-mouse/human CD207 (4C7)BioLegendBarcode: CGATTTGTATTCCCTTotalSeq-A0438 anti-mouse/rat KKC2BioLegendBarcode: GAGCTTGTACCGCTTTotalSeq-A0439 anti-mouse CD201 (RCR-16)BioLegend141509; RRID: AB_2800655TotalSeq-A0440 anti-mouse CD169 (Siglec-1) (N1/12)BioLegend142425; RRID: AB_2783106TotalSeq-A0441 anti-mouse CD71 (3D6.112)BioLegend113824; RRID: AB_2800574TotalSeq-A0442 anti-mouse Notch (HMN1-12)BioLegend130617; RRID: AB_2783085TotalSeq-A0443 anti-mouse CD41 (MWReg30)BioLegend133937; RRID: AB_2800635TotalSeq-A0444 anti-mouse CXCR4 (L276F12)BioLegend146520; RRID: AB_2800682TotalSeq-A0445 anti-human TMEM119 (A16075D)BioLegend853303; RRID: AB_2801201TotalSeq-A0446 anti-human CD93 (VIMD2)BioLegend336121; RRID: AB_2750379TotalSeq-A0448 anti-mouse CD204 (Msr1) (1F8C33)BioLegend154703; RRID: AB_2783126TotalSeq-A0449 anti-mouse CD326 (Ep-CAM) (G8.8)BioLegend118237; RRID: AB_2800586TotalSeq-A0450 anti-mouse IgM (RMM-1)BioLegend406535; RRID: AB_2783322TotalSeq-A0551 anti-mouse CD301a (MGL1) (LOM-8.7)BioLegend145611; RRID: AB_2783114TotalSeq-A0552 anti-mouse CD304 (Neuropilin-1) (3E12)BioLegend145215; RRID: AB_2750383TotalSeq-A0554 anti-mouse CD309 (VEGFR2, Flk-1) (89B3A5)BioLegend121921; RRID: AB_2783066TotalSeq-A0555 anti-mouse CD36 (HM36)BioLegend102621; RRID: AB_2800557TotalSeq-A0556 anti-mouse CD370 (CLEC9A-DNGR1) (7H11)BioLegendBarcode: AACTCAGTTGTGCCGTotalSeq-A0557 anti-mouse CD38 (90)BioLegend102733; RRID: AB_2750556TotalSeq-A0558 anti-mouse CD55 (DAF) ( RIKO-3)BioLegend131809; RRID: AB_2783086TotalSeq-A0559 anti-mouse CD63 (NVG-2)BioLegend143915; RRID: AB_2783109TotalSeq-A0560 anti-mouse CD68 (FA-11)BioLegend137031; RRID: AB_2783099TotalSeq-A0561 anti-mouse CD79b (Igβ) (HM79-12)BioLegend132811; RRID: AB_2783087TotalSeq-A0562 anti-mouse CD83 (Michel-19)BioLegend121519; RRID: AB_2783061TotalSeq-A0563 anti-mouse CX3CR1 (SA011F11)BioLegend149041; RRID: AB_2783121TotalSeq-A0564 anti-mouse Folate Receptor β (FR-β) (10/FR2)BioLegend153307; RRID: AB_2800690TotalSeq-A0565 anti-mouse MERTK (Mer) (2B10C42)BioLegendBarcode: AGTAGAGCAACTCGTTotalSeq-A0566 anti-mouse CD301b (MGL2) (URA-1)BioLegend146817; RRID: AB_2783115TotalSeq-A0567 anti-mouse Tim-4 (RMT4-54)BioLegend130011; RRID: AB_2783084TotalSeq-A0568 anti-mouse/rat XCR1 (ZET)BioLegend148227; RRID: AB_2783120TotalSeq-A0569 anti-human CD338 (5D3)BioLegend332021; RRID: AB_2783216TotalSeq-A0570 anti-mouse/rat CD29 (HMβ1-1)BioLegend102233; RRID: AB_2783042TotalSeq-A0571 anti-mouse IgD (11-26c.2a)BioLegend405745; RRID: AB_2783321TotalSeq-A0572 anti-human C5L2 (1D(-M12)BioLegend342407; RRID: 2783226TotalSeq-A0573 anti-mouse CD140a (APA5)BioLegend135917; RRID: AB_2783094TotalSeq-A0574 anti-human CD235a (HI264)BioLegend349117; RRID: AB_2783242TotalSeq-A0575 anti-human CD49a (TS2/7)BioLegend328315; RRID: AB_2783195TotalSeq-A0576 anti-human CD49d (9F10)BioLegend304337; RRID: AB_2783166TotalSeq-A0577 anti-human CD73 (AD2)BioLegend344029; RRID: AB_2783228TotalSeq-A0578 anti-human CD79aBioLegendBarcode: CTTATCACCCGCTTTTotalSeq-A0579 anti-human CD9 (H19a)BioLegend312119; RRID: AB_2783182TotalSeq-A0580 anti-human mast cell tryptaseBioLegendBarcode: ACTGATAGACCCGCTTotalSeq-A0581 anti-human TCR (3C10)BioLegend351733; RRID: AB_2783246TotalSeq-A0582 anti-human TCR (B6)BioLegend331433; RRID: AB_2800863TotalSeq-A0583 anti-human TCR (B3)BioLegend331311; RRID: AB_2783207TotalSeq-A0584 anti-human TCR (6B11)BioLegend342923; RRID: AB_2783227TotalSeq-A0586 anti-human CD354 (Trem-26)BioLegend314910;TotalSeq-A0588 anti-human CD202b (HIT2)BioLegend334213; RRID: AB_2810511TotalSeq-A0590 anti-human CD305 (NKTA255)BioLegend342805; RRID: AB_2800911TotalSeq-A0591 anti-human LOX1 (15C4)BioLegend358611; RRID: AB_2800987TotalSeq-A0592 anti-human CD158b (DX27)BioLegend312615; RRID: AB_2800818TotalSeq-A0593 anti-human CD203c (NP4D6)BioLegend324627; RRID: AB_2800849TotalSeq-A0595 anti-mouse CD11a (M17/4)BioLegend101125; RRID: AB_2783036TotalSeq-A0596 anti-mouse ESAM (1G8/ESAM)BioLegend136209; RRID: AB_2800642TotalSeq-A0597 anti-human CD209 (9E9A8)BioLegend330119; RRID: AB_2783206TotalSeq-A0599 anti-human CD158e1 (DX9)BioLegend312723; RRID: AB_2800819TotalSeq-A0600 anti-human CD158f (UP-R1)BioLegend341307; RRID: AB_2800904TotalSeq-A0801 anti-human CD337 (P30-15)BioLegend325221; RRID: AB_2800852TotalSeq-A0803 anti-human CD253 (RIK-2)BioLegend308211; RRID: AB_2800803TotalSeq-A0804 anti-human CD186 (K041E5)BioLegend356021; RRID: AB_2800961TotalSeq-A0807 anti-mouse CD200R (OX-110)BioLegend123913; RRID: AB_2800594TotalSeq-A0808 anti-mouse CD193 (J073E5)BioLegend144523; RRID: AB_2800673TotalSeq-A0809 anti-mouse CD200R3 (Ba13)BioLegend142209; RRID: AB_2800657TotalSeq-A0810 anti-mouse CD138 (281-2)BioLegend142532; RRID: AB_2800658TotalSeq-A0811 anti-mouse CD317 (27)BioLegend127027; RRID: AB_2800623TotalSeq-A0812 anti-mouse CD105 (MJ7/18)BioLegend120421; RRID: AB_2800587TotalSeq-A0813 anti-mouse CD9 (MZ3)BioLegend124819; RRID: AB_2800600TotalSeq-A0814 anti-human CD205 (HD30)BioLegend342211; RRID: AB_2800908TotalSeq-A0816 anti-human CD271 (ME20.4)BioLegend345123; RRID: AB_2814273TotalSeq-A0817 anti-human CD109 (W7C5)BioLegend323307; RRID: AB_2800848TotalSeq-A0819 anti-human CD126 (UV4)BioLegend352813; RRID: AB_2800939TotalSeq-A0821 anti-human CD164 (67D2)BioLegend324809; RRID: AB_2800850TotalSeq-A0822 anti-human CD142 (NY2)BioLegend365207; RRID: AB_2801007TotalSeq-A0824 anti-mouse P2X7R (1F11)BioLegend148711; RRID: AB_2800683TotalSeq-A0825 anti-mouse CD371 (5D3/Clec12a)BioLegend143407; RRID: AB_2800668TotalSeq-A0826 anti-human CD307c (H5/FcRL3)BioLegend374411; RRID: AB_2801022TotalSeq-A0827 anti-mouse CD22 (OX-97)BioLegend126113; RRID: AB_2800614TotalSeq-A0828 anti-human CD307d (413D12)BioLegend340209; RRID: AB_2800902TotalSeq-A0829 anti-human CD307e (509f6)BioLegend340307; RRID: AB_2800903TotalSeq-A0830 anti-human CD319 (162.1)BioLegend331821; RRID: AB_2800872TotalSeq-A0831 anti-human CD138 (DL-101)BioLegend352325; RRID: AB_2800938TotalSeq-A0834 anti-mouse CD39 (Duha59)BioLegend143813; RRID: AB_2800669TotalSeq-A0835 anti-mouse CD314 (NKG2D) (CX5)BioLegend130215; RRID: AB_2814023TotalSeq-A0836 anti-mouse DR3 (4C12)BioLegend144413; RRID: AB_2814048TotalSeq-A0837 anti-mouse IL33RA (DIH9)BioLegend145317; RRID: AB_2800680TotalSeq-A0839 anti-mouse Ly49H (3D10)BioLegend144715; RRID: AB_2814049TotalSeq-A0841 anti-mouse Ly49D (4E5)BioLegend138309; RRID: AB_2800647TotalSeq-A0843 anti-human CD199 (L053E8)BioLegend358919; RRID: AB_2810569TotalSeq-A0844 anti-human CD45RB (MEM-55)BioLegend310209; RRID: AB_2810471TotalSeq-A0845 anti-human CD99 (3B2/TA8)BioLegend371317: RRID: AB_2801017TotalSeq-A0846 anti-mouse CD185 (L138D7)BioLegend145535; RRID: AB_2800681TotalSeq-A0848 anti-mouse TIGIT (1G9)BioLegend142115; RRID: AB_2800656TotalSeq-A0849 anti-mouse CD80 (16-10A1)BioLegend104745; RRID: AB_2813935TotalSeq-A0850 anti-mouse CD49a (Hmα1)BioLegend142613; RRID: AB_2800659TotalSeq-A0851 anti-mouse CD1d (1B1)BioLegend123529; RRID: AB_2800593TotalSeq-A0852 anti-mouse CD226 (10E5)BioLegend128823; RRID: AB_2810393TotalSeq-A0853 anti-human CD371 (50C1)BioLegend353613; RRID: AB_2800948TotalSeq-A0857 anti-mouse CD34 (HM34)BioLegend128619; RRID: AB_2810392TotalSeq-A0858 anti-human CD46 (TRA-2-10)BioLegend352415; RRID: AB_2810557TotalSeq-A0861 anti-human CD151 (50-6)BioLegend350409; RRID: AB_2810554TotalSeq-A0862 anti-human CD218a (H44)BioLegend313815; RRID: AB_2810476TotalSeq-A0863 anti-human CD257 (1D6)BioLegend366509; RRID: AB_2810575TotalSeq-A0866 anti-human CLEC1B (AYP1)BioLegend372009; RRID: AB_2814333TotalSeq-A0867 anti-human CD94 (DX22)BioLegend305521; RRID: AB_2814142TotalSeq-A0868 anti-human IgE (MHE-18)BioLegend325517; RRID: AB_2814186TotalSeq-A0869 anti-human CD365 (1D12)BioLegend353907; RRID: AB_2814294TotalSeq-A0870 anti-human CD150 (A12)BioLegend306313; RRID: AB_2814146TotalSeq-A0871 anti-human CD162 (KPL-1)BioLegend328821; RRID: AB_2814192TotalSeq-A0875 anti-mouse TLR4 (MTS510)BioLegend117614; RRID: AB_2810352TotalSeq-A0876 anti-mouse CD300c_d (TX52)BioLegend148005; RRID: AB_2810413TotalSeq-A0877 anti-mouse JAML (4E+10)BioLegend128507; RRID: AB_2810391TotalSeq-A0881 anti-mouse CD272 (6A6)BioLegend139113; RRID: AB_2814041TotalSeq-A0882 anti-mouse PIRA_PIRB (6C1)BioLegend144105; RRID: AB_2810412TotalSeq-A0883 anti-mouse CD26 (H194-112)BioLegend137811; RRID: AB_2810405TotalSeq-A0884 anti-mouse DLL1 (HMD1-3)BioLegend128315; RRID: AB_2810390TotalSeq-A0885 anti-mouse CD270 (HMHC-1B18)BioLegend136307; RRID: AB_2810403TotalSeq-A0890 anti-mouse 4_1BB (TKS-1)BioLegend107109: RRID: AB_2813955TotalSeq-A0891 anti-mouse ENPP (YE1/19.1)BioLegend149209; RRID: AB_2814052TotalSeq-A0892 anti-mouse CD2 (RM2-5)BioLegend100117; RRID: AB_2810312TotalSeq-A0894 anti-human Ig (MHL-38)BioLegend316531; RRID: AB_2810479TotalSeq-A0895 anti-mouse/human Mac2 (LM3/38)BioLegend125421; RRID: AB_281384TotalSeq-A0896 anti-human CD85j (GHI/75)BioLegend333723; RRID: AB_2814225TotalSeq-A0897 anti-human CD23 (EBVCS-5)BioLegend338523; RRID: AB_2814235TotalSeq-A0898 anti-human Ig (MHK-49)BioLegend316627; RRID: AB_2814170TotalSeq-A0899 anti-human HLA-A2 (BB7.2)BioLegend343331; RRID: AB_2810540TotalSeq-A0900 anti-human CD198 (L263G8)BioLegend360607; RRID: AB_2810572TotalSeq-A0901 anti-human GARP (7B11)BioLegend352515; RRID: AB_2814283TotalSeq-A0904 anti-mouse CD31 (390)BioLegend102437; RRID: AB_2810335TotalSeq-A0905 anti-mouse CD107a (1D4B)BioLegend121635; RRID: AB_2810369TotalSeq-A0916 anti-mouse CD124 (I015F8)BioLegend144809; RRID: AB_2814050TotalSeq-A0917 anti-mouse CD95 (SA367H8)BioLegend152614; RRID: AB_2810418TotalSeq-A0923 anti-human NKp80 (5D12)BioLegend346709; RRID: AB_2814274TotalSeq-A5106 anti-mouse Ly6DBioLegendBarcode: ATGTCCTACCTCAAAChemicals, peptides, and recombinantproteinsAntigenfixDiapathP0014β-mercaptoethanolSigma-AldrichM3148Calcium chloride dihydrateMerck1023821000Collagenase ASigma-Aldrich11088793001D-(-)-FructoseMerck-Millipore1040071000D(+)-SaccharoseVWR InternationalPROL27483.294DAPIInvitrogenD1306; RIDD: AB_2629482DMEMInvitrogen41965-039Dnase ISigma-Aldrich04 536 282 001Donkey SerumAbcamab7475EDTAWestburg51234EGTASigma-AldrichE3889EosinVWR InternationalMERC 1.15935FcBlock 2.4G2BiocerosN/AFCSBodinco5010Fixable Viability due Live/Dead - eFluor506eBioscience65-0866-18Fixable Viability due Live/Dead - eFluor780eBioscience65-0865-18Fixation/Permeabilization Solution KitBD Cytofix/Cytoperm554714FoxP3 Transcription factor staining buffer kiteBioscience00-5523-00GentamicinGibco15710-049GlutaMAXThermo Fisher35050-038Gluteraldehyde 25%Sigma-AldrichG5882Goat SerumSigma-AldrichG9023HematoxylinVWR InternationalMERC1.05174HEPESSigma-AldrichH3375IsopropanolVelT0108MethanolMerck Millipore13680502Paraformaldehyde 10%EMS15712Phenol RedSigma-AldrichP3532ProLong DiamondThermo FisherP36970Rat SerumSigma-AldrichR9759Ribonucleoside Vanadyl ComplexesSigma-AldrichR3380-5MLRoche Protector RNAse inhibitorSigma-Aldrich3335399001RPMI 1640Gibco52400-025SaponinSigma-Aldrich452110X SDSSigma-Aldrich71736-100MLSodium bicarbonateSigma-Aldrich792519Sodium chlorideSigma-Aldrich746398Sodium Dihydrogen Phosphate MonohydrateSigma-Aldrich1063461000Sodium phosphate dibasic dihydrateSigma-Aldrich71643Tissue-Tek O.C.TSakura Finetek4583UltraPure™ Salmon Sperm DNA SolutionThermo Fisher Scientific15632011XyleneProlaboPROL28973.363Critical commercial assaysALLin HS Red Taq Mastermix 2xhighQuHQ.HSM0350RNEasy Plus Micro KitQIAGEN74034SensiFAST cDNA Synthesis KitBiolineBIO-65054SensiFAST SYBR No-ROX KitBiolineBIO-98020Visium Spatial Gene Expression Slide and Reagent Kit10X Genomics1000184Oligonucleotides:qPCRFlt3 – qPCR FWDIDTGTGACTGGCCCCCTGGATAACGAGFlt3 – qPCR REVIDTTCCAAGGGCGGGTGTAACTGAACTXcr1 - qPCR FWDIDTAGAGACACCGAACAGTCAGGCTXcr1 - qPCR REVIDTTGTCCAGTTGCTGAAGGCTCTCCd209a - qPCR FWDIDTGCACTCCATCAAAGGCTTTGGCCd209a - qPCR REVIDTCAAACAGCTAGGAAGAGCACCTGCcr7 - qPCR FWDIDTAGAGGCTCAAGACCATGACGGACcr7 - qPCR REVIDTTCCAGGACTTGGCTTCGCTGTAMafb - qPCR FWDIDTTGAATTTGCTGGCACTGCTGMafb - qPCR REVIDTAAGCACCATGCGGTTCATACACd5l - qPCR FWDIDTGAGGACACATGGATGGAATGTCd5l - qPCR REVIDTACCCTTGTGTAGCACCTCCACd207 - qPCR FWDIDTCCGAAGCGCACTTCACAGTCd207 - qPCR REVIDTGCAGATACAGAGAGGTTTCCTCASpp1 - qPCR FWDIDTCCATCTCAGAAGCAGAATCTCCTTSpp1 - qPCR REVIDTGGTCATGGCTTTCATTGGAATTGata6 - qPCR FWDIDTCAGCAGGACCCTTCGAAACGata6 - qPCR REVIDTCCATTCATCTTGCTGTAGAGACCGpnmb - qPCR FWDIDTAGCACAACCAATTACGTGGCGpnmb - qPCR REVIDTCTTCCCAGGAGTCCTTCCAPla2g7 - qPCR FWDIDTCTTCAAGCCCTTAGTGAGGACCPla2g7 - qPCR REVIDTTGCGATGTCCTTTGGAGTCTGGCd9 - qPCR FWDIDTGCTACTCGAGCCATGCCGGTCAAAGGAGGTAGCCd9 - qPCR REVIDTCACTTGGTACCGACCATTTCTCGGCTCCTGCGCd36 - qPCR FWDIDTGGAGCCATCTTTGAGCCTTCACd36 - qPCR REVIDTGAACCAAACTGAGGAATGGATCTIl1b - qPCR FWDIDTGCAACTGTTCCTGAACTCAACTIl1b - qPCR REVIDTATCTTTTGGGGTCCGTCAACTTnf - qPCR FWDIDTTCTTCTCATTCCTGCTTGTGGTnf - qPCR REVIDTGGTCTGGGCCATAGAACTGAIl10 - qPCR FWDIDTAAGGCAGTGGAGCAGGTGAAIl10 - qPCR REVIDTCCAGCAGACTCAATACACACIl18 - qPCR FWDIDTACTGTACAACCGCAGTAATACGCIl18 - qPCR REVIDTAGTGAACATTACAGATTTATCCCActb - qPCR FWDIDTGCTTCTAGGCGGACTGTTACTGAActb - qPCR REVIDTGCCATGCCAATGTTGTCTCTTATOligonucleotides:Molecular cartography probesFLT3Resolve Bio.
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PMC8809252
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
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P0C6WPDGFRBResolve Bio.
|
PMC8809252
|
Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.
|
P0D6VLGR5Resolve Bio.
|
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